Navigation Links
The Use of MasterTaq Polymerase for PCR with Humic Material-Contaminated ,,, DNA

PCR amplification. Two universal bacterial primers EUB008 and EUB1492 were used to amplify 16S rDNA genes from the extracted and purified
chromosomal DNA.11,12 PCR was performed with a Eppendorf Mastercycler gradient** as follows: 50 pmol of each primer, 2.5 mol of each deoxyribonucleoside triphosphate, 300 g bovine serum albumin (BSA), 1x reaction buffer, 1x TaqMaster PCR enhancer and 1U MasterTaq DNA Polymerase were adjusted to a final volume of 100 l with sterile water. From 80 ng to 500 ng of template DNA was added to the reaction mixture after preheating to 70C to avoid nonspecific annealing of the primers to nontarget DNA. The cycles used were:

1 cycle at 70C for 1 min, followed by 38 cycles at 95C for 1 min, 40C for 1 min, 72C for 3 min and 1 final cycle at 95C for 1 min, 40C for 1 min and at 72C for 10 min. In parallel, the same experiment was performed using a conventional Taq polymerase, which was routinely used in our lab and considered to be of maximal suitability for PCR. DNA extracted from E. coli was used as a positive control.

The use of high concentrations of BSA in the hot-start PCR was shown to decrease the sensitivity of the reaction against components in the soil extracts and to make PCR product formation reproducible.


Fig. 1: Use of MasterTaq and conventional Taq polymerase for the amplification of 16S rDNA genes from humic-acid contaminated DNA extracted from a marine sediment. PCR products were analyzed on a 1% agarose gel.

Lanes 1 and 4: E. coli-DNA (= positive controls)
Lanes 2 and 5: negative control (H2O)
Lanes 3 and 6: community DNA, extracted from a marine sediment



'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. Amplification of the amoA Gene of Nitrosococcus oceani Bacteria Using Eppendorf MasterTaq
2. Amplification of Mouse cDNAs for Microarrays Using the Eppendorf MasterTaq Kit
3. Enhancing PCR* Efficiency with MasterTaq
4. Novel PCR Enhancing Factor Improves Performance of Pfu DNA Polymerase
5. Pfu DNA Polymerase as a Hot Start Alternative
6. PCR Performance Comparisons Between pfuturbo and Taq DNA Polymerases
7. pfuturbo DNA Polymerase: A High-Performance, High-Fidelity Enzyme Ideal for PCR Cloning
8. Unique DNA Polymerase Formulation Excels in a Broad Range of PCR Applications
9. Enhanced Amplification of Long Targets with PfuTurbo DNA Polymerase
10. High-Fidelity PCR with a Novel Polymerase Mixture
11. Optimizing pfuturbo DNA Polymerase Amplification Reactions with Perfect Match PCR Enhancer
Post Your Comments:
(Date:8/21/2014)... MD (PRWEB) August 21, 2014 DNA ... making progress in its development of JRC-LAMP-vax, an innovative ... cedar pollen. Phase IC studies began this month in ... to hay fever. , The Japanese red cedar releases ... people in Japan. Mountain Cedar pollen wreaks similar havoc ...
(Date:8/21/2014)... , August 21, 2014 ... http://www.researchandmarkets.com/research/3zh3q8/global_hormone ) has announced the addition of ...  report to their offering.       ... as hormone therapy, is a means of ... produced by the body. This type of ...
(Date:8/21/2014)... , Aug. 21, 2014  Decision Resources Group ... Brazil and Mexico ... of several newly approved agents, the anticipated label ... and the launch of emerging biologics will substantially ... particular, the emerging IL-5 inhibitors will introduce a ...
(Date:8/21/2014)... , Aug. 21, 2014  Four-dimensional (4-D) printing ... based on external stimuli such as changes in ... a disruptive effect in multiple industries.  In the ... landscape by printing objects ranging from human organs ... sectors. Logo - http://photos.prnewswire.com/prnh/20140820/138289 ...
Breaking Biology Technology:Relief for Allergy Sufferers Finally in Sight: Immunomic Therapeutics Closes in on Treatment for Hay Fever in Japan, U.S. 2Relief for Allergy Sufferers Finally in Sight: Immunomic Therapeutics Closes in on Treatment for Hay Fever in Japan, U.S. 3Relief for Allergy Sufferers Finally in Sight: Immunomic Therapeutics Closes in on Treatment for Hay Fever in Japan, U.S. 4Global Hormone Replacement Therapy Market 2014-2018: Key Vendors are Actavis, Bayer, Merck & Co, Novartis and Novo Nordisk 2Asthma Agents Currently Face a Suboptimal Coverage Environment in Brazil and Mexico, and as the Treatment Armamentarium Expands, Competitiveness Will Further Intensify 2Asthma Agents Currently Face a Suboptimal Coverage Environment in Brazil and Mexico, and as the Treatment Armamentarium Expands, Competitiveness Will Further Intensify 3Frost & Sullivan: 4-D Printing to Usher in Age of Low-Labor, Fast-Paced Product Manufacturing 2Frost & Sullivan: 4-D Printing to Usher in Age of Low-Labor, Fast-Paced Product Manufacturing 3Frost & Sullivan: 4-D Printing to Usher in Age of Low-Labor, Fast-Paced Product Manufacturing 4
... VisEn Medical Inc., a leader in fluorescence in vivo ... of its new Cat-K FAST (TM) ... with disease progression and therapeutic response in vivo . ... therapeutic target in a range of bone-related diseases, including cancer metastasis ...
... 7 Mylan Inc. (Nasdaq: MYL ) today ... from the U.S. Food and Drug Administration (FDA) for its ... , , Bicalutamide Tablets are the generic version of ... of approximately $322 million for the 12 months ending March ...
... MATEO, Calif., July 7 King Pharmaceuticals, Inc. (NYSE: ... today announced that on July 2, 2009, King met with the ... Letter regarding the New Drug Application (NDA) for REMOXY(R). The ... to resubmit the REMOXY(R) NDA and to address all FDA comments ...
Cached Biology Technology:VisEn Launches New Cat-K FAST(TM) Fluorescence Molecular Imaging Agent 2King and Pain Therapeutics Announce REMOXY(R) NDA Update 2King and Pain Therapeutics Announce REMOXY(R) NDA Update 3
(Date:8/21/2014)... unknown process for harvesting energy and producing oxygen from ... University scientist has discovered. The discovery lays the foundation ... energy from the Sun, and understanding dense blooms like ... worldwide. A paper describing the discovery will be published ... journal Science on 21 August 2014. , ...
(Date:8/21/2014)... aimed at improving the food intake, health and quality ... is among three projects at the University of Waterloo ... Canadian Institutes of Health Research (CIHR). , Professor ... at Waterloo, and a Schlegel research chair in nutrition ... investigate why many Canadians living in long-term care homes ...
(Date:8/21/2014)... Ever heard of the water window? It consists of ... are not absorbed by the water in biological tissues. ... develop coherent radiations within the water window. These could ... a high-contrast image of the biological samples or to ... study identifies the physical mechanism needed to efficiently generate ...
Breaking Biology News(10 mins):Hot-spring bacteria reveal ability to use far-red light for photosynthesis 2Hot-spring bacteria reveal ability to use far-red light for photosynthesis 3Grants will fund landmark aging research at Waterloo 2Water window imaging opportunity 2
... to be satisfied? Less than half as much as ... snacking study. Using chocolate chips, apple pie, and potato ... Wansink designed a study to determine if people who ... hungrier or satisfied fifteen minutes after eating., ,Two groups ...
... in the American Chemical Society,s (ACS,) award-winning Global ... to improve the sensitivity of the test often used to ... possessions of passengers at airport security screening stations. Based ... ACS, The Journal of Physical Chemistry C , the ...
... Aviv Prof. Nathan Nelson of Tel Aviv University,s ... Center has been awarded the 2012 Israel Prize in ... awarded in Jerusalem on the eve of Israel,s Independence ... and molecular biology, Prof. Nelson is internationally renowned for ...
Cached Biology News:New American Chemical Society podcast: Boosting the sensitivity of airport security screening 2
Practical tool for easy, effective sealing of 0.2ml PCR tubes and plates using cap strips....
... by BioLegend, the Quansys Biosciences Q-Plex Array ... of cytokines/chemokines for human, mouse, and rat ... a fully quantitative ELISA-based test where 4 ... each well of a 96-well plate in ...
The strip cap tool is a used for properly seating 8- and 12-cap strips in microplates or strip tubes. A grooved channel on one side is designed to seat domed caps, while the flat edge on the opposite...
... The expertise that made the original Gene Pulser ... system available has been applied in the Gene ... The modular design of the Gene Pulser II ... accessory that has the widest range of settings ...
Biology Products: